Preparation and Evaluation of a Self-Nanoemulsifying Drug Delivery System Loaded with Heparin Phospholipid Complex

A self-nanoemulsifying drug delivery system (SNEDDS) was developed to enhance the absorption of heparin after oral administration, in which heparin was compounded with phospholipids to achieve better fat solubility in the form of heparin-phospholipid (HEP-Pc) complex. HEP-Pc complex was prepared using the solvent evaporation method, which increased the solubility of heparin in n-octanol. The successful preparation of HEP-Pc complex was confirmed by differential scanning calorimetry (DSC), Fourier-transform infrared (FT-IR) spectroscopy, NMR, and SEM. A heparin lipid microemulsion (HEP-LM) was prepared by high-pressure homogenization and characterized. HEP-LM can enhance the absorption of heparin after oral administration, significantly prolong activated partial thromboplastin time (APTT) and thrombin time (TT) in mice, and reduce fibrinogen (FIB) content. All these outcomes indicate that HEP-LM has great potential as an oral heparin formulation.     

Polysaccharide‐based artificial extracellular matrix: Preparation and characterization of three‐dimensional,macroporous chitosan,and heparin composite scaffold

Scaffold‐guided tissue engineering based on synthetic and natural occurring polymers has gained many interests in recent year. In this study, the development of a chitosan‐heparin artificial extracellular matrix (AECM) is reported. Three‐dimensional, macroporous composite AECMs composed of heparin (Hep) and chitosan (Chito) were prepared by an interpolyelectrolyte complex/lyophilization method. The Chito‐Hep composite AECMs were, respectively, crosslinked with glutaraldehyde, as well as cocrosslinked with N,N‐(3‐dimethylaminopropyl)‐N′‐ethyl carbodiimide (EDC/NHS) and N‐hydroxysuccinimide (NHS). The crosslinking reactions were examined by FT‐IR analysis. In physiological buffer solution (PBS), the EDC/NHS‐crosslinked Chito‐Hep composite AECM showed a relative lower water retention ratio than its glutaraldehyde‐crosslinked counterparts. The EDC/NHS‐crosslinked Chito‐Hep composite AECMs showed excellent biocompatibility, according to the results of the in vitro cytotoxic test. This result suggested that the EDC/NHS‐crosslinked Chito‐Hep composite AECMs might be a potential biomaterial for scaffold‐guided tissue engineering applications. © 2008 Wiley Periodicals, Inc. J Appl Polym Sci, 2008     

Immobilization of biologically active species on PA‐6 foils treated by a dielectric barrier discharge

In the field of biomaterials and biomedical devices, surface activation has been focused on creating functional groups capable of preferential adsorption of biologically active species (proteins, enzymes, cells, drugs, etc.). In this way an interface can be created between the synthetic material and the biological medium, with the aim of increasing the compatibility of the implant with the human organism. In our experiments a dielectric barrier discharge (DBD), in helium at atmospheric pressure, was used as the source of energy capable of creating active centers that render the functionalized surface favorable to immobilization of biological molecules. Retention of immunoglobulin (IgG) and heparin biomolecules on polyamide‐6 (PA‐6) surfaces after treatment by the DBD was analyzed by atomic force microscopy, adhesion evaluation, and measurement of the contact angle titration in order to assess this incorporation on the treated surfaces. The marked adsorption of the biomolecules on the active sites created by DBD on the exposed surfaces also was related to a complex set of processes, such as enhanced roughness, increased surface wettability, and modified distribution of cationic and anionic groups on the treated surfaces. All these factors could promote interfacial interactions between the specific groups of the biomolecules existing in the biological medium and the type of cationic and/or anionic groups present on the surface. The efficiency of the DBD treatment showed that the DBD technique is useful for preactivation of the polymer surface for immobilization of other biologically active species (such as drugs and enzymes). © 2003 Wiley Periodicals, Inc. J Appl Polym Sci 90: 1985–1990, 2003     

Effect of heparin on high glucose induced proliferation and expression of matrix metalloproteinases in normal human mesangial cells

Background The pathogenesis of diabetic nephropathy (DN) is a complex pathophysiological process. Its precise mechanism is not fully known. In recent years it has been recognized that synthesis of various extracelluar matrix (ECM) components may increase, and that degradation of ECM may decrease in DN. It was reported heparin could inhibit mesangial cells proliferation in vitro. The main aim of this study is to explore whether heparin inhibits proliferation of mesangial cells grown in high glucose concentration and to measure the effect of heparin on matrix metalloproteinases (MMPs) expression in mesangial cells. Methods The medium contained either low glucose (5 mmol/L) or high glucose (25 mmol/L). The concentrations of heparin in the culture medium were 0, 25, 50, 100, 200 or 400 μg/mL. A metabolic (WST-1) assay was used to measure mesangial cell proliferation and Western blot analysis was used to measure MMPs expression of mesangial cells. Results Normal human mesangial cell (NHMC) proliferation was higher in high glucose (HG) medium than in low glucose (LG) medium. They showed a 1.93 fold expansion after 72 h in high glucose in contrast to a 1.63 fold expansion in low glucose. In the presence of heparin, mesangial cells proliferation was inhibited, which was more obvious at high glucose concentrations than at low glucose concentrations. In high glucose, with heparin concentration of 50, 100, 200 and 400 μg/mL, the mesangial cells showed a 0.61 fold, 0.52 fold, 0.52 fold and 0.41 fold reductions in cell number compared to cells grown without heparin. In low glucose, only concentrations of 200 μg/mL and 400 μg/mL showed reduction in cell number, namely 0.54 fold and 0.45 fold, when compared to cells grown without heparin. In Western blot analysis, MMP₁, MMP₂, MMP₃ and MMP₉ was expressed by mesangial cells expressed in both high and low glucose concentrations, which was more prominent in high glucose medium. Incubation of heparin further increased expression of MMP₁, MMP₂, MMP₃ and MMP₉. Conclusions This study suggests that glucose can accelerate mesangial cell proliferation while heparin can reduce proliferation, being more obvious at high glucose concentrations. Higher glucose concentrations led to increased MMP expression, which may take part in the regulation of mesangial matrix synthesis and degradation. Addition of heparin resulted in a corresponding increase in MMP expression, most notably at high glucose concentrations, indicating a potentially renoprotective role in DN. Foundation item: Project (30370663) supported by the National Natural Science Foundation of China     

PET膜的接枝改性及其血液相容性研究

对PET(聚对苯二甲酸乙二醇酯)材料进行表面改性,以提高材料表面亲水性和抗凝血性能,采取紫外共辐照方法和逐步偶合接枝方法,先在PET表面接枝聚乙二醇,然后通过化学偶合方法在聚乙二醇末端接枝抗凝血药物肝素。改性后PET材料的表面亲水性和抗凝血性能得到很大改善。PET表面亲水性随着接枝PEG的分子量和反应浓度的增大而增强。使用亲水性高聚物PEG作为空间臂,可以很好地保持肝素的生物活性。通过紫外改性成功地在材料表面接枝了PEG和肝素,很好地改善了材料的生物相容性。     

Synthesis and Cell Growth Inhibitory Activity of Six Non-glycosaminoglycan-Type Heparin-Analogue Trisaccharides

The design and synthesis of heparin mimetics with high anticancer activity but no anticoagulant activity is an important task in medicinal chemistry. Herein, we present the efficient synthesis of five Glc-GlcA-Glc-sequenced and one Glc-IdoA-Glc-sequenced non-glycosaminoglycan, heparin-related trisaccharides with various sulfation/sulfonylation and methylation patterns. The cell growth inhibitory effects of the compounds were tested against four cancerous human cell lines and two non-cancerous cell lines. Two d -glucuronate-containing tetra-O-sulfated, partially methylated trisaccharides displayed remarkable and selective inhibitory effects on the growth of ovary carcinoma (A2780) and melanoma (WM35) cells. Methyl substituents on the glucuronide unit proved to be detrimental, whereas acetyl substituents were beneficial to the cytostatic activity of the sulfated derivatives.     

Pellets for oral administration of low-molecular-weight heparin

Background: Oral absorption of low-molecular-weight heparin (LMWH) is limited by its molecular size and negative charge. It has been shown previously that orally administered polymeric nano- or microparticles containing encapsulated LMWH have led to gastrointestinal absorption of heparin in rabbits. Method: Based on these investigations, pellets containing two LMWHs, enoxaparin (MW 4500 Da) or bemiparin (MW 3600 Da), and Eudragit®RS30D (ERS), were prepared using extrusion/ spheronization technique. Uncoated or coated (ERS) pellets were evaluated in vitro and in vivo on rabbits. Results: Enoxaparin pellets showed fast in vitro release in phosphate buffer (pH 7.4) and prolonged in vivo drug absorption after a single oral dose of 600 anti-Xa IU/ kg of body weight, leading to relative bioavailabilities ranging from 9.7 ± 1.9% to 12.8 ± 2.7% and anti-Xa activity over the curative dose. Bemiparin included in matrix pellets of ERS and coated with ERS exhibited in vitro prolonged release up to 4 hours and in vivo anti-Xa activity below the therapeutic minimum value of 0.1 IU/mL. Conclusion: This study presents LMWH in a pellet dosage form, which compared to nano- or microparticles, may offer a more convenient and industrializable way of manufacture leading to an easier scale-up process.     

细菌纤维素/肝素复合物的制备与力学性能

采用浸泡和交联法将肝素(Hep)固定在细菌纤维素(BC)上,研究了BC/Hep复合物的结构与力学性能。傅立叶变换红外光谱结果表明,BC结构中出现了Hep的特征吸收峰;扫描电子显微镜和X射线衍射显示,浸泡或交联后,在Hep的作用下,BC的结构变得紧密,结晶度下降,晶粒尺寸增大,其中交联法制备的复合物的结构与结晶性能变化更为明显。拉伸性能实验表明,与纯BC相比,浸泡和交联制备的BC/Hep复合物力学性能增强,交联法制备的复合物力学性能优于浸泡法,Hep溶液浓度为1.5 g/100 m L时其拉伸强度最大,可达到177.82 MPa,与纯BC相比提高了76.3%。     

Clinical and Molecular Characterization of Patients with Mucopolysaccharidosis Type I in an Algerian Series

Mucopolysaccharidoses (MPS’s) represent a subgroup of lysosomal storage diseases related to a deficiency of enzymes that catalyze glycosaminoglycans degradation. Mucopolysaccharidosis type I (MPS I) is a rare autosomal recessive disorder caused by a deficiency of α-l-iduronidase encoded by the IDUA gene. Partially degraded heparan sulfate and dermatan sulfate accumulate progressively and lead to multiorgan dysfunction and damage. The aim of this study is to describe the clinical, biochemical, and molecular characteristics of 13 Algerian patients from 11 distinct families. MPS I diagnosis was confirmed by molecular study of the patients’ IDUA gene. Clinical features at the diagnosis and during the follow-up are reported. Eighty-four percent of the studied patients presented with a mild clinical phenotype. Molecular study of the IDUA gene allowed the characterization of four pathological variations at the homozygous or compound heterozygote status: IDUA NM_00203.4:c.1598C>G-p.(Pro533Arg) in 21/26 alleles, IDUA NM_00203.4:c.532G>A-p.(Glu178Lys) in 2/26 alleles, IDUA NM_00203.4:c.501C>G-p.(Tyr167*) in 2/26 alleles, and IDUA NM_00203. 4: c.1743C>G-p.(Tyr581*) in 1/26 alleles. This molecular study unveils the predominance of p.(Pro533Arg) variation in our MPS I patients. In this series, the occurrence of some clinical features linked to the Scheie syndrome is consistent with the literature, such as systematic valvulopathies, corneal opacity, and umbilical hernia; however, storage signs, facial dysmorphic features, and hepatomegaly were more frequent in our series. Screening measures for these debilitating diseases in highly consanguineous at-risk populations must be considered a priority health problem.     

High-Elastic Blood Compatible Material

Abstract

Materials with unique complex of the properties of elasticity and tensile strength processed from the natural rubber latex (NRL) were modified to improve their blood compatibility. ESCA, ATR-IR, UV-spectroscopy and other methods were used for appraisement of the grade of modification, structure and properties of material. Thromboresistancy was studied in in vivo and ex vivo tests. It was shown that blood compatibility is raised after thorough purification of the material from non-rubber components by means of two-stage extraction changing the physical-chemical parameters of the surface. Two different principles were used in the process of modification: 1) coating by thin polyurethane (PU) coverings with improved thromboresistant properties-thus the problem of providing high adhesion interaction of covering with the latex base was solved; 2) heparin surface immobilization-higher efficiency of modification of latex material in gel-form without preliminary protein adsorption was shown. Modification allows to increase blood compatibility of the latex materials preserving at the same time their elasticity and tensile strength.